To compare a single-step medium with a sequential medium on human blastocyst development rates, aneuploidy rates, and clinical outcomes.
Retrospective cohort study of IVF cycles that used Sage advantage sequential medium (n = 347) and uninterrupted Sage 1-step medium (n = 519) from July 1, 2016, to December 31, 2017, in an academic fertility center. Main outcome measures are blastocyst formation rates per two-pronuclear (2PN) oocyte and aneuploidy rates per biopsy.
Of all IVF cycles, single-step medium yielded higher blastocyst formation rate (51.7% vs 43.4%) but higher aneuploidy rate (54.0% vs 45.8%) compared with sequential medium. When stratified by maternal age, women under age 38 had no difference in blastocyst formation (52.2% vs 50.2%) but a higher aneuploidy rate (44.5% vs 36.4%) resulting in a lower number of euploid blastocysts per cycle (2.6 vs 3.3) when using single-step medium compared to sequential medium. In cycles used single-step medium, patients ≥ age 38 had higher blastocyst rate (48.0% vs 33.6%), but no difference in aneuploidy rate (68.8% vs 66.0%) or number of euploid embryos (0.8 vs 1.1). For patients reaching euploid embryo transfer, there was no difference in clinical pregnancy rates, miscarriage rates, or live birth rates between two culture media systems.
Our study demonstrates an increase in aneuploidy in young women whose embryos were cultured in a single-step medium compared to sequential medium. This study highlights the importance of culture conditions on embryo ploidy and the need to stratify by patient age when examining the impact of culture conditions on overall cycle potential.
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Machtinger R, Racowsky C. Chapter 12: Culture systems: single step. In: Smith G, editor. Embryo culture: methods and protocols, methods in molecular biology: Springer Science+Business Media, LLC; 2012.
Swain JE. Controversies in ART: considerations and risks for uninterrupted embryo culture. Reprod BioMed Online. 2019;39(1):19–26. https://doi.org/10.1016/j.rbmo.2019.02.009.
Macklon NS, Pieters MH, Hassan MA, Jeucken PH, Eijkemans MJ, Fauser BC. A prospective randomized comparison of sequential versus monoculture systems for in-vitro human blastocyst development. Hum Reprod. 2002;17(10):2700–5.
Paternot G, Debrock S, D'Hooghe TM, Spiessens C. Early embryo development in a sequential versus single medium: a randomized study. Reprod Biol Endocrinol. 2010;8:83.
Campo R, Binda MM, Van Kerkhoven G, Frederickx V, Serneels A, Roziers P, et al. Critical reappraisal of embryo quality as a predictive parameter for pregnancy outcome: a pilot study. Facts Views Vis Obgyn. 2010;2(4):289–95.
Ciray HN, Aksoy T, Goktas C, Ozturk B, Bahceci M. Time-lapse evaluation of human embryo development in single versus sequential culture media--a sibling oocyte study. J Assist Reprod Genet. 2012;29(9):891–900. https://doi.org/10.1007/s10815-012-9818-7.
Reed ML, Hamic A, Thompson DJ, Caperton CL. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study. Fertil Steril. 2009;92(5):1783–6. https://doi.org/10.1016/j.fertnstert.2009.05.008.
Summers MC, Bird S, Mirzai FM, Thornhill A, Biggers JD. Human preimplantation embryo development in vitro: a morphological assessment of sibling zygotes cultured in a single medium or in sequential media. Hum Fertil (Camb). 2013;16(4):278–85. https://doi.org/10.3109/14647273.2013.806823.
Hardarson T, Bungum M, Conaghan J, Meintjes M, Chantilis SJ, Molnar L, et al. Noninferiority, randomized, controlled trial comparing embryo development using media developed for sequential or undisturbed culture in a time-lapse setup. Fertil Steril. 2015;104(6):1452-9.e1-4. https://doi.org/10.1016/j.fertnstert.2015.08.037.
Basile N, Morbeck D, García-Velasco J, Bronet F, Meseguer M. Type of culture media does not affect embryo kinetics: a time-lapse analysis of sibling oocytes. Hum Reprod. 2013;28(3):634–41. https://doi.org/10.1093/humrep/des462.
Sfontouris IA, Kolibianakis EM, Lainas GT, Petsas GK, Tarlatzis BC, Lainas TG. Blastocyst development in a single medium compared to sequential media: a prospective study with sibling oocytes. Reprod Sci. 2017;24(9):1312–8. https://doi.org/10.1177/1933719116687653.
Sepúlveda S, Garcia J, Arriaga E, Diaz J, Noriega-Portella L, Noriega-Hoces L. In vitro development and pregnancy outcomes for human embryos cultured in either a single medium or in a sequential media system. Fertil Steril. 2009;91(5):1765–70. https://doi.org/10.1016/j.fertnstert.2008.02.169.
Werner MD, Hong KH, Franasiak JM, Forman EJ, Reda CV, Molinaro TA, et al. Sequential versus Monophasic Media Impact Trial (SuMMIT): a paired randomized controlled trial comparing a sequential media system to a monophasic medium. Fertil Steril. 2016;105(5):1215–21. https://doi.org/10.1016/j.fertnstert.2016.01.005.
Sfontouris IA, Martins WP, Nastri CO, Viana IG, Navarro PA, Raine-Fenning N, et al. Blastocyst culture using single versus sequential media in clinical IVF: a systematic review and meta-analysis of randomized controlled trials. J Assist Reprod Genet. 2016;33(10):1261–72. https://doi.org/10.1007/s10815-016-0774-5.
Swain JE. Controversies in ART: can the IVF laboratory influence preimplantation embryo aneuploidy? Reprod BioMed Online. 2019. https://doi.org/10.1016/j.rbmo.2019.06.009.
Cimadomo D, Scarica C, Maggiulli R, Orlando G, Soscia D, Albricci L, et al. Continuous embryo culture elicits higher blastulation but similar cumulative delivery rates than sequential: a large prospective study. J Assist Reprod Genet. 2018;35(7):1329–38. https://doi.org/10.1007/s10815-018-1195-4.
Munné S, Alikani M, Ribustello L, Colls P, Martínez-Ortiz PA, McCulloh DH, et al. Euploidy rates in donor egg cycles significantly differ between fertility centers. Hum Reprod. 2017;32(4):743–9. https://doi.org/10.1093/humrep/dex031.
Franasiak JM, Forman EJ, Hong KH, Werner MD, Upham KM, Treff NR, et al. The nature of aneuploidy with increasing age of the female partner: a review of 15,169 consecutive trophectoderm biopsies evaluated with comprehensive chromosomal screening. Fertil Steril. 2014;101(3):656–63.e1. https://doi.org/10.1016/j.fertnstert.2013.11.004.
Thomas MR, Sparks AE, Ryan GL, Van Voorhis BJ. Clinical predictors of human blastocyst formation and pregnancy after extended embryo culture and transfer. Fertil Steril. 2010;94(2):543–8. https://doi.org/10.1016/j.fertnstert.2009.03.051.
Gardner DK, Lane M, Stevens J, Schlenker T, Schoolcraft WB. Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer. Fertil Steril. 2000;73(6):1155–8.
Babariya D, Fragouli E, Alfarawati S, Spath K, Wells D. The incidence and origin of segmental aneuploidy in human oocytes and preimplantation embryos. Hum Reprod. 2017;32(12):2549–60. https://doi.org/10.1093/humrep/dex324.
Vera-Rodríguez M, Michel CE, Mercader A, Bladon AJ, Rodrigo L, Kokocinski F, et al. Distribution patterns of segmental aneuploidies in human blastocysts identified by next-generation sequencing. Fertil Steril. 2016;105(4):1047-55.e2. https://doi.org/10.1016/j.fertnstert.2015.12.022.
Munné S, Chen S, Colls P, Garrisi J, Zheng X, Cekleniak N, et al. Maternal age, morphology, development and chromosome abnormalities in over 6000 cleavage-stage embryos. Reprod BioMed Online. 2007;14(5):628–34.
Picton HM, Elder K, Houghton FD, Hawkhead JA, Rutherford AJ, Hogg JE, et al. Association between amino acid turnover and chromosome aneuploidy during human preimplantation embryo development in vitro. Mol Hum Reprod. 2010;16(8):557–69. https://doi.org/10.1093/molehr/gaq040.
Liang B, Gao Y, Xu J, Song Y, Xuan L. Shi T, et al. Fertil Steril: Raman profiling of embryo culture medium to identify aneuploid and euploid embryos; 2019. https://doi.org/10.1016/j.fertnstert.2018.11.036.
Kropp J, Salih SM, Khatib H. Expression of microRNAs in bovine and human pre-implantation embryo culture media. Front Genet. 2014;5:91. https://doi.org/10.3389/fgene.2014.00091.
Suh N, Blelloch R. Small RNAs in early mammalian development: from gametes to gastrulation. Development. 2011;138(9):1653–61. https://doi.org/10.1242/dev.056234.
Machtinger R, Racowsky C. Culture systems: single step. Methods Mol Biol. 2012;912:199–209. https://doi.org/10.1007/978-1-61779-971-6_12.
Brison DR, Houghton FD, Falconer D, Roberts SA, Hawkhead J, Humpherson PG, et al. Identification of viable embryos in IVF by non-invasive measurement of amino acid turnover. Hum Reprod. 2004;19(10):2319–24. https://doi.org/10.1093/humrep/deh409.
Kaing A, Kroener LL, Tassin R, Li M, Liu L, Buyalos R, et al. Earlier day of blastocyst development is predictive of embryonic euploidy across all ages: essential data for physician decision-making and counseling patients. J Assist Reprod Genet. 2018;35(1):119–25. https://doi.org/10.1007/s10815-017-1038-8.
Tiegs AW, Sun L, Patounakis G, Scott RT. Worth the wait? Day 7 blastocysts have lower euploidy rates but similar sustained implantation rates as Day 5 and Day 6 blastocysts. Hum Reprod. 2019;34(9):1632–9. https://doi.org/10.1093/humrep/dez138.
The authors thank the staffs at Stanford IVF center for their clinical support of the study.
J.D. and Q.Z. were involved with study design, execution, and statistical analysis. J.D. wrote the manuscript. C.C. and R.K. coordinated data collection and statistical analysis. R.L. contributed interpretation of the data and edit of the manuscript. B.B. devised, supervised the study and edited the manuscript. All authors reviewed the manuscript and provided critical feedback and discussion.
Conflict of interest
Both Cengiz Cinnioglu and Refik Kayali disclosed that they are employees of Igenomix Inc. All other authors do not have conflicts of interest to disclose.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
The work was done in Stanford Medicine Fertility and Reproductive Health Services.
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Deng, J., Zhao, Q., Cinnioglu, C. et al. The impact of culture conditions on blastocyst formation and aneuploidy rates: a comparison between single-step and sequential media in a large academic practice. J Assist Reprod Genet (2020). https://doi.org/10.1007/s10815-019-01621-8
- Culture media
- Embryo development